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Supreme Catalytic Properties of Enzyme Nanoparticles Based on Ferritin Self-Assembly
ACS Applied Bio Materials ( IF 4.6 ) Pub Date : 2020-09-08 , DOI: 10.1021/acsabm.0c00961
Qingmei Chen 1, 2, 3 , Dong Men 4 , Tianyu Sun 1 , Yan Zhang 1 , Qianqian Yuan 1 , Taidou Hu 1 , Zhangsheng Hu 1 , Jun Wu 1, 2, 3 , Yiqun Deng 1, 2, 3 , Xian-En Zhang 5 , Jikai Wen 1, 2, 3
Affiliation  

In this study, we aimed to address if enzymes self-assembled on the biological nanosphere matrix would have some nanoscale effects and then improve the catalytic ability and other characteristics especially for the resistance to adverse conditions. Initially, we evaluated two different forms of ferritins, DNA-binding protein from starved cells (Dps) and recombinant heavy chain of human ferritin (rHF), fused with methyl parathion hydrolase (MPH) to be self-assembled as the dodecamer and 24-mer enzyme nanoparticles (enzyme-NPs), Dps–MPH and MPH–rHF. Both self-assembled enzyme-NPs showed superior catalytic activity and kcat drastically increased, but MPH–rHF showed better performance than Dps–MPH. Next, Escherichia coli alkaline phosphatase (EAP) and the canonical mutant EAP(D101S) were chosen to confirm if rHF-based self-assembly generally improves the catalysis of enzymes with the different catalytic abilities. Additionally, restoration of the enzyme native forms on the rHF shell by increasing the flexibility of the polylinker between the catalytic units and the nanoscaffold further improved the catalytic activities of enzyme-NPs. Some of the nanoparticles exhibited better residual activities under extreme conditions. Conclusively, the enzyme-NPs based on rHF self-assembly provided an effective strategy for enzyme engineering.

中文翻译:

基于铁蛋白自组装的酶纳米粒子的最高催化性能

在这项研究中,我们旨在解决在生物纳米球基质上自组装的酶是否会产生一些纳米级效应,然后提高催化能力和其他特性,特别是对不利条件的抵抗力。最初,我们评估了两种不同形式的铁蛋白,来自饥饿细胞的 DNA 结合蛋白 (Dps) 和重组人铁蛋白重链 (rHF),与甲基对硫磷水解酶 (MPH) 融合以作为十二聚体和 24- mer 酶纳米颗粒 (enzyme-NPs)、Dps-MPH 和 MPH-rHF。两种自组装酶-NPs都表现出优异的催化活性,k cat急剧增加,但MPH-rHF表现出比Dps-MPH更好的性能。接下来,大肠杆菌选择碱性磷酸酶 (EAP) 和典型突变体 EAP (D101S) 来确认基于 rHF 的自组装是否普遍提高了具有不同催化能力的酶的催化作用。此外,通过增加催化单元和纳米支架之间的多接头的柔韧性来恢复rHF壳上的酶天然形式,进一步提高了酶-NPs的催化活性。一些纳米粒子在极端条件下表现出更好的残留活性。总之,基于rHF自组装的酶-NPs为酶工程提供了一种有效的策略。
更新日期:2020-10-21
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